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05-745R-S Sigma-AldrichAnti-trimethyl-Histone H3 (Lys4) Antibody, clone 15-10C-E4, Trial Size

Use Anti-trimethyl-Histone H3 (Lys4) Antibody, clone 15-10C-E4, Recombinant, Trial Size (rabbit monoclonal antibody) proven in WB, ChIP, ChIP-seq, ELISA, PIA, Mplex, DB to detect trimethyl-Histone H3 (Lys4) also known as H3K4me3.

MSDS (material safety data sheet) or SDS, CoA and CoQ, dossiers, brochures and other available documents.

Research Category: Epigenetics & Nuclear Function Research Sub-Category: Histones Gene Symbol: H3F3A, H3F3B UniProt Number: Q16695

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Key Spec Table

Species Reactivity	Key Applications	Host	Format	Antibody Type
H	WB, ChIP, ChIP-seq, DB, ELISA, PIA, Mplex	Rb	Purified	Monoclonal Antibody

Description
Catalogue Number	05-745R-S
Replaces	05-745
Trade Name	Upstate
Description	Anti-trimethyl-Histone H3 (Lys4) Antibody, clone 15-10C-E4, Trial Size
Overview	Recombinant version of Cat. # 05-745, clone MC315
Alternate Names	H3K4me3 Histone H3 (tri methyl K4)
Background Information	Histone H3 is one of the five main histone proteins involved in the structure of chromatin in eukaryotic cells. Featuring a main globular domain and a long N-terminal tail, H3 is involved with the structure of the nucleosomes of the 'beads on a string' structure. The N-terminal tail of histone H3 protrudes from the globular nucleosome core and can undergo several different types of epigenetic modifications that influence cellular processes. These modifications include the covalent attachment of methyl or acetyl groups to lysine and arginine amino acids and the phosphorylation of serine or threonine.

References

Product Information
Format	Purified
Presentation	Protein A purified, recombinant antibody in 60 mM Tris, 90 mM NaCl, 0.03% NaN3, 60 mM Glycine, pH 7.5, 40% Glycerol.
Quality Level	MQ100

Applications
Application	Use Anti-trimethyl-Histone H3 (Lys4) Antibody, clone 15-10C-E4, Recombinant, Trial Size (rabbit monoclonal antibody) proven in WB, ChIP, ChIP-seq, ELISA, PIA, Mplex, DB to detect trimethyl-Histone H3 (Lys4) also known as H3K4me3.
Key Applications	Western Blotting Chromatin Immunoprecipitation (ChIP) ChIP-seq Dot Blot ELISA Peptide Inhibition Assay Multiplexing
Application Notes	ChIP-seq Analysis: Representative lot data. Chromatin immunoprecipitation was performed using the Magna ChIP™ HiSens kit (cat# 17-10460), 3 µL of Anti-trimethyl-Histone H3 (Lys4) antibody (cat# 05-745R-S), 20 µL Protein A/G beads, and 1e6 crosslinked HeLa cell chromatin followed by DNA purification using magnetic beads. Libraries were prepared from Input and ChIP DNA samples using standard protocols with Illumina barcoded adapters, and analyzed on Illumina HiSeq instrument. An excess of eighteen million reads from FastQ files were mapped using Bowtie (http://bowtie-bio.sourceforge.net/manual.shtml) following TagDust (http://genome.gsc.riken.jp/osc/english/dataresource/) tag removal. Peaks were identified using MACS (http://luelab.dfci.harvard.edu/MACS/), with peaks and reads visualized as a custom track in UCSC Genome Browser (http://genome.ucsc.edu) from BigWig and BED files. The highest 25% of peaks identified in the 05-745R and 07-473 datasets showed 99% overlap with peaks identified in the ENCODE H3K4me3 BROAD Histone track for HeLa S3.

Biological Information
Immunogen	BSA-conjugated, synthetic peptide containing the sequence …RTme3KQT… in which me3K corresponds to trimethyl-lysine 4 of human histone H3.
Clone	15-10C-E4
Host	Rabbit
Specificity	Recognizes Histone H3 containing trimethyl-lysine 4 and, to a lesser extent, dimethyl-lysine 4. See certificate of analysis for further details.
Isotype	IgG
Species Reactivity	Human
Species Reactivity Note	Broad species cross-reactivity expected due to sequence homology
Antibody Type	Monoclonal Antibody
Entrez Gene Number	NM_003493
Entrez Gene Summary	Histones are basic nuclear proteins that are responsible for the nucleosome structure of the chromosomal fiber in eukaryotes. Nucleosomes consist of approximately 146 bp of DNA wrapped around a histone octamer composed of pairs of each of the four core histones (H2A, H2B, H3, and H4). The chromatin fiber is further compacted through the interaction of a linker histone, H1, with the DNA between the nucleosomes to form higher order chromatin structures.
Gene Symbol	H3F3A H3F3B
Modifications	Methylation
UniProt Number	Q16695
UniProt Summary	FUNCTION: SwissProt: Q16695 # Core component of nucleosome. Nucleosomes wrap and compact DNA into chromatin, limiting DNA accessibility to the cellular machineries which require DNA as a template. Histones thereby play a central role in transcription regulation, DNA repair, DNA replication and chromosomal stability. DNA accessibility is regulated via a complex set of post-translational modifications of histones, also called histone code, and nucleosome remodeling. SIZE: 136 amino acids; 15508 Da SUBUNIT: The nucleosome is a histone octamer containing two molecules each of H2A, H2B, H3 and H4 assembled in one H3-H4 heterotetramer and two H2A-H2B heterodimers. The octamer wraps approximately 147 bp of DNA. SUBCELLULAR LOCATION: Nucleus. TISSUE SPECIFICITY: Expressed in testicular cells. DEVELOPMENTAL STAGE: Expressed during S phase, then expression strongly decreases as cell division slows down during the process of differentiation. PTM: Acetylation is generally linked to gene activation. Acetylation on Lys-10 impairs methylation at Arg-9. Acetylation on Lys-19 and Lys-24 favors methylation at Arg-18 (By similarity). & Citrullination at Arg-9 and/or Arg-18 by PADI4 impairs methylation and represses transcription (By similarity). & Asymmetric dimethylation at Arg-18 by CARM1 is linked to gene activation. Symmetric dimethylation at Arg-9 by PRMT5 is linked to gene repression (By similarity). & Methylation at Lys-5, Lys-37 and Lys-80 are linked to gene activation. Methylation at Lys-5 facilitates subsequent acetylation of H3 and H4. Methylation at Lys-80 is associated with DNA double-strand break (DSB) responses and is a specific target for TP53BP1. Methylation at Lys-10 and Lys-28 are linked to gene repression. Methylation at Lys-10 is a specific target for HP1 proteins (CBX1, CBX3 and CBX5) and prevents subsequent phosphorylation at Ser-11 and acetylation of H3 and H4. Methylation at Lys-5 and Lys-80 require preliminary monoubiquitination of H2B at 'Lys-120'. Methylation at Lys-10 and Lys-28 are enriched in inactive X chromosome chromatin (By similarity). & Phosphorylated at Thr-4 by GSG2/haspin during prophase and dephosphorylated during anaphase. At centromeres, specifically phosphorylated at Thr-12 from prophase to early anaphase. Phosphorylated at Ser-11 during the whole mitosis. Phosphorylation at Ser-11, which is linked to gene activation, prevents methylation at Lys-10 but facilitates acetylation of H3 and H4. Phosphorylated at Ser-29 by MLTK isoform 1, RPS6KA5 or AURKB during mitosis or upon ultraviolet B irradiation (By similarity). & Phosphorylation at 'Ser-11' is crucial for chromosome condensation and cell-cycle progression during mitosis and meiosis. In addition phosphorylation at 'Ser-11' is important during interphase because it enables the transcription of genes following external stimulation, like stress or growth factors. Phosphorylation at 'Ser-11' is also an essential regulatory mechanism for neoplastic cell transformation. Phosphorylation at 'Ser-11' by AURKB/Aurora-B mediates the dissociation of HP1 proteins (CBX1, CBX3 and CBX5) from heterochromatin. & Ubiquitinated (By similarity). SIMILARITY: SwissProt: Q16695 ## Belongs to the histone H3 family.
Molecular Weight	17 kDa

Physicochemical Information

Dimensions

Materials Information

Toxicological Information

Safety Information according to GHS

Safety Information

Product Usage Statements
Quality Assurance	Routinely evaluated by immunoblot on HeLa nuclear extract
Usage Statement	Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.

Storage and Shipping Information
Storage Conditions	Stable for 1 year at -20°C from date of receipt. Handling Recommendations: Upon receipt, and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance. Note: Variability in freezer temperatures below -20°C may cause glycerol-containing solutions to become frozen during storage.

Storage and Shipping Information

Storage Conditions

Stable for 1 year at -20°C from date of receipt.
Handling Recommendations: Upon receipt, and prior to removing the cap, centrifuge the vial and gently mix the solution. Aliquot into microcentrifuge tubes and store at -20°C. Avoid repeated freeze/thaw cycles, which may damage IgG and affect product performance. Note: Variability in freezer temperatures below -20°C may cause glycerol-containing solutions to become frozen during storage.

Packaging Information
Material Size	10 µL

Transport Information

Supplemental Information

Specifications

Global Trade Item Number
Catalogue Number	GTIN
05-745R-S	04053252946844

Documentation

Anti-trimethyl-Histone H3 (Lys4) Antibody, clone 15-10C-E4, Trial Size SDS

Title
Safety Data Sheet (SDS)

Anti-trimethyl-Histone H3 (Lys4) Antibody, clone 15-10C-E4, Trial Size Certificates of Analysis

Title	Lot Number
Anti-trimethyl-Histone H3 (Lys4), clone 15-10C-E4, Recombinant, Trial Size - Q2378155	Q2378155
Anti-trimethyl-Histone H3 (Lys4), clone 15-10C-E4, Recombinant, Trial Size - 3191456	3191456
Anti-trimethyl-Histone H3 (Lys4), clone 15-10C-E4, Recombinant, Trial Size - 3234578	3234578
Anti-trimethyl-Histone H3 (Lys4), clone 15-10C-E4, Recombinant, Trial Size - 3819942	3819942
Anti-trimethyl-Histone H3 (Lys4), clone 15-10C-E4, Recombinant, Trial Size - 3845335	3845335
Anti-trimethyl-Histone H3 (Lys4), clone 15-10C-E4, Recombinant, Trial Size Monoclonal Antibody	3059114
Anti-trimethyl-Histone H3 (Lys4), clone 15-10C-E4, Recombinant, Trial Size Monoclonal Antibody	3111605

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